A number of organophosphorus (“OP”) compounds used by the agriculture industry and the military are highly toxic and thus hazardous to human health and harmful to the environment. For example, acetylcholinesterase-inhibiting OP compounds comprise the active ingredient of pesticides such as paraoxon as well as G-type nerve agents such as Sarin and Soman, etc., developed for chemical warfare. Thus, it is very important to be able to detoxify such OP compounds and to decontaminate surfaces and substances contaminated with these compounds.
One approach being investigated as a potential solution to this problem is enzyme-mediated decontamination. For example, a class of enzymes known as organophosphorus acid (“OPA”) anhydrolases (“OPAA”) (EC 3.1.8.2) can catalyze the hydrolysis of a variety of OP compounds, including pesticides and fluorinated “G-type” nerve agents. These anhydrolases are mass produced via overexpression within recombinant organisms as described by U.S. Pat. No. 5,928,927 to Cheng et al., which is incorporated herein by reference.
One of the organophosphorus compounds, (cyclohexyl methylphosphonofluoridate, CAS registry number 329-99-7, known as GF, is very toxic to humans. The median lethal dose (LD50) for humans is estimated to be about 0.35 g/man when contact is through skin. The estimated LCt50 for inhalation is estimated to be 70 mg min/m3. No efficient and easily produced catalyst for GF degradation in the environment or in vivo is known. The native OPAA enzyme has been described to possess catalytic activity against various chemical nerve agents, but its activity against the particularly toxic agent GF (cyclohexyl methylphosphonofluoridate) is limited, and therefore, is marginally useful as a decontaminant or as a medical countermeasure for GF poisoning.
Efforts on producing organophosphorus acid anhydrolases for detoxifying organophosphorus compounds are well known in the art.
U.S. Pat. No. 5,928,927 to Cheng et al. teaches expression and composition comprising wild-type organophosphorus acid anhydrolases (“OPAA-2”) from the bacteria strain Alteromonas sp. JD6.5.
U.S. 2013/0071394 to Troyer et al. teaches compositions and combinations containing an organophosphorus bioscavenger and a hyaluronan-degrading enzyme that can be used to treat or prevent organophosphorus poisoning, including nerve agent poisoning and pesticide poisoning. However, the bioscavenger that Troyer utilizes is a wild-type OPAA.
Similar to '394, U.S. Pat. No. 8,920,824 to Rosenberg teaches treating humans exposed to sarin by inhalation of wild-type OPAA.
U.S. Pat. No. 9,017,982 to Shah et al. teaches a non-wild-type organophosphorus acid anhydrolases having an amino acid substitution at position 212, such that the mutated OPAA may degrade (ethyl {2-[bis(propan-2-yl)amino]ethyl}sulfanyl) (methyl)phosphinate and other V-agents. However, the mutation occurs only at position 212 and the reported increase in catalytic activity is on V-type agents.
U.S. 2015/0017186 to Troyer et al. teaches a composition comprising an organophosphorus bioscavenger and a hyaluronan-degrading enzyme, and its use thereof to treat organophosphorus poisoning.
U.S. 2016/0355792 to Pegan teaches a mutated OPAA having mutation at positions Y212F, V342L, and I215Y for degrading VX and VR. However, the reported increase in catalytic activity is only for V-type agents.
U.S. Pat. No. 9,512,413 to Hansen, et al. teaches an organophosphorus hydrolase from Ciona savignyl or Ciona intestinalis, having at least one mutation in positions 212 to 314 for removing organophosphorus compounds.
U.S. Pat. No. 9,771,566 to Hansen, et al. teaches an organophosphorus hydrolase variant, having mutations at one or more of the following positions: 34, 37, 38, 58, 59, 61, 63, 91, 94, 96, 111, 164, 165, 166, 169, 170, 171, 193, 194, 216, 219, 243, 245, 246, 247, 248, 250, 266, 290, 291, 293, and 312. However, Hansen does not teach that one or more of these mutations of the hydrolase is to hydrolyze GF.
Therefore, new compounds and methods to effectively detoxify GF are needed.